The development of science and the resulting increase in human life expectancy have led to increased problems of various diseases, aging, and organ damage due to various accidents, and thus there is a growing interest in the tissue-engineering to regenerate tissues.
A cell carrier (Scaffold) refers to a physical scaffold and adhesion substrate made possible for culture in vitro and implantation of tissue cells, and these cell carriers are used for cell transplantation for tissue regeneration. In the case of epithelial cells used for skin tissue regeneration, it is required to be used after incubation in the cell carrier because adhesion of the cells and accompanying transfer and proliferation are possible only when there is a contact area between the substrates.
Since such cell carrier for skin tissue regeneration is directly inserted into the body, collagen and the like, which are derived from terrestrial animals such as pigs and cattle, which are biodegradable, are currently used. However, the use of collagen derived from terrestrial animals has been avoided due to safety issues such as mad cow disease and foot-and-mouth disease, and interest in natural substances derived from safer marine organisms has been increasing.
The oceans are known to account for more than 70% of the surface area of the Earth's surface and to be the source of hundreds of thousands of species of biological materials, which account for 80% of whole animals in the Earth. Recently, marine natural substances obtained from marine include a large amount of various minerals and physiologically active materials and have proved that they have excellent biocompatibility compared to other animal natural products. Thus, in advanced countries, marine organisms are noticed as new substances for physiological activity and new materials for the tissue-engineering. Thus, in order to explore new natural materials and to create new added value, the biotechnology and bioorganic chemistry techniques are applied to the marine natural materials to concentrate on development of biomaterials
In particular, chitin and chitosan in marine natural materials refer to natural cationic polysaccharides present in the cell walls of crustacean shells, insect epidermis, and some microorganisms. Chitosan, prepared by alkaline deacetylation of chitin, is composed of β(1→4)-bond N-acetyl-D-glucosamine units and has a variety of biological activities. Chitooligosaccharide is hydrolyzed to chitosan derivatives. Unlike chitosan, chitooligosaccharide has free amino groups with a shorter chain length in the D-glucosamine unit. For such reason, it is known that chitooligosaccharide is soluble in natural aqueous solutions and thus can be easily applied to in vitro and in vivo systems and has anti-tumor, anti-fungal, antibacterial, anti-virus and reactive oxygen species (ROS) scavenging ability according to a range of their molecular weight (kDa). However, there is no hitherto-known method of utilizing chitooligosaccharide for cell carrier for skin tissue regeneration depending on a range of the molecular weight (kDa).
As a prior art of the present invention, a method for manufacturing a carrier of transplanting pancreatic islet cells using atelocollagen, and artificial pancreas manufactured by the same method is disclosed in Korea Patent Registration No. 10-1327630, but relates to a carrier of transplanting pancreatic islet cells in which alginate and atelocollagen derived from cationized pigs are used as constituents to increase the survival rate of islet cells and insulin secretion. Also, nanoparticles for therapeutic gene delivery and method for preparing the same are disclosed in Korea Patent Publication No. 10-2009-0097582, which relates to a gene medicine carrier which has improved biocompatibility and stability by cross-linking chitosan to alginate. In addition, the preparation method of bilayer scaffold in one-step and tissue regeneration method of bilayer scaffold obtained thereby are disclosed in Korea Patent Registration No. 10-1333381, but relates to a method of preparing a scaffold by cross-linking biocompatibility materials such as gelatin and polyvinyl alcohol with EDC.
Thus, in the patent documents as described above, there is no known cell carrier for skin tissue regeneration in which chitooligosaccharide (COS) having different molecular weights (kD) from each other is cross-linked to fish-derived collagen (Fish collagen, FC) and alginate (A) scaffold to improve the biocompatibility of fibroblasts.